The evolutionary analysis of the Tnt1 retrotransposon in Nicotiana species reveals the high variability of its regulatory sequences

We studied the evolution of the tobacco Tnt1 retrotransposon by analyzing Tnt1 partial sequences containing both coding domains and U3 regulatory sequences obtained from a number of Nicotiana species. We detected three different subfamilies of Tnt1 elements, Tnt1A, Tnt1B, and Tnt1C, that differ completely in their U3 regions but share conserved flanking coding and LTR regions. U3 divergence between the three subfamilies is found in the region that contains the regulatory sequences that control the expression of the well-characterized Tnt1-94 element. This suggests that expression of the three Tnt1 subfamilies might be differently regulated. The three Tnt1 subfamilies were present in the Nicotiana genome at the time of species divergence, but have evolved independently since then in the different genomes. Each Tnt1 subfamily seems to have conserved its ability to transpose in a limited and different number of Nicotiana species. Our results illustrate the high variability of Tnt1 regulatory sequences. We propose that this high sequence variability could allow these elements to evolve regulatory mechanisms in order to optimize their coexistence with their host genome.      

Agrobacterium rhizogenes—transformed plant roots as a source of grapevine viruses for purification

Agrobacterium rhizogenes-mediated transformation was applied toVitis spp. andNicotiana spp. infected by different grapevine phloem-limited viruses (grapevine fleck virus, grapevine virus A, grapevine virus B) to obtain root cultures for virus purification. All plant species were successfully transformed, and several clones were established in liquid culture. Transformed grapevine roots contained as much virus as non transformed roots and more than leaves, as assessed by ELISA and thin sectioning. Likewise, transformed roots ofNicotiana benthamiana Domin. contained in average more GVA than leaves, especially those at the base and the top of the plant, whereas withNicotiana occidentalis wheel., GVB was apparently less concentrated than in leaves.Nicotiana root grew faster than those ofVitis. All viruses multiplied and persisted in root cultures, which were successfully used for purification. Virus yields were the same (GFkV and GVB) or higher (GVB) than those reported in the literature. Grapevine roots may prove useful for culturing and purifying other non-mechanically transmissible grapevine viruses.     

Mitochondrial growth and division during the cell cycle in HeLa cells

The growth and division of mitochondria during the cell cycle was investigated by a morphometric analysis of electron micrographs of synchronized HeLa cells. The ratio of total outer membrane contour length to cytoplasmic area did not vary significantly during the cell cycle, implying a continuous growth of the mitochondrial outer membrane. The mean fraction of cytoplasmic area occupied by mitochondrial profiles was likewise found to remain constant, indicating that the increase in total mitochondrial volume per cell occurs continuously during interphase, in such a way that the mitochondrial complement occupies a constant fraction( approximately 10-11(percent)) of the volume of the cytoplasm. The mean area, outer membrane contour length, and axis ratio of the mitochondrial profiles also did not vary appreciably during the cell cycle; furthermore, the close similarity of the frequency distributions of these parameters for the six experimental time-points suggested a stable mitochondrial shape distribution. The constancy of both the mean mitochondrial profile area and the number of mitochondrial profiles per unit of cytoplasmic area was interpreted to indicate the continuous division of mitochondria at the level of the cell population. Furthermore, no evidence was found for the occurrence of synchronous mitochondrial growth and division within individual cells. Thus, it appears that, in HeLa cells, there is no fixed temporal relationship between the growth and division of mitochondria and the events of the cell cycle. A number of statistical methods were developed for the purpose of making numerical estimates of certain three-dimensional cellular and mitochondrial parameters. Mean cellular and cytoplasmic volumes were calculated for the six time-points; both exhibited a nonlinear, approx. twofold increase. A comparison of the axis ratio distributions of the mitochondrial profiles with theoretical distributions expected from random sectioning of bodies of various three-dimensional shapes allowed the derivation of an "average" mitochondrial shape. This, in turn, permitted calculations to be made which expressed the two-dimensional results in three-dimensional terms. Thus, the estimated values for the number of mitochondria per unit of cytoplasmic volume and for the mean mitochondrial volume were found to remain constant during the cell cycle, while the estimated number of mitochondria per cell increase approx. twofold in an essentially continuous manner.     

Microtubules and cyclic amp in human leukocytes: on the order of things

We have shown previously that the β-adrenergic agonist isoproterenol (2μM) and the phosphodiesterase inhibitor isobutylmethylxanthine (1 mM) produce a much greater increase in cyclic AMP in human leukocytes that have been pretreated with colchicine (or with other agents that affect microtubule assembly) than in control leukocytes. The effects of colchicines were both time- and dose-dependant. These and other data suggested that the generation of cyclic AMP is normally restricted by an intact system of cytoplasmic microtubules. If so, then the same time and dose dependencies might apply to other colchicines-induced changes in leukocyte function. We have now assayed the distribution of concanavalin A (Con A)-receptor complexes on the leukocyte membrane, taking into account that leukocytes competent to assemble microtubules show a uniform distribution of surface- bound Con A whereas microtubule-deficient cells accumulate Con A in surface caps. We have found that the effect of colchicine on capping is also both time- and dose dependent, and that the dose-response relationships conform to those required to increase cyclic AMP levels. These findings provide further evidence that both colchicine-induced Con-A capping and colchicine- induced cyclic AMP generation depend upon the relaxation of constraints normally imposed by cytoplasmic microtubules upon the plasma membrane, which limit, respectively, lateral mobility of the lectin-receptor complexes, and expression of hormone-sensitive adenylate cyclase. Moreover, colchicine-induced Con-A cap formation is not affected even by very large changes in leukocyte cyclic AMP levels. Thus, elevated cyclic AMP levels do not appear to promote the dissolution of microtubules; rather, the dissolution of microtubules permits the generation of increased amounts of cyclic AMP.     

Monitoring of airborne pollen and pollen calendar of Cosenza,Southern Italy

Summary From May 1986 to May 1989 surveys of airborne biological particles have been performed in the atmosphere of Cosenza, Italy with the aim of monitoring the presence of airborne pollen. The survey station is situated at Arcavacata of Rende, a hilly area 474 meters above sea level, 7 km north-west of the main town. The sampler (VPPS 2000) is located about 20 meters above ground level. The monitoring, performed in accordance with the criteria suggested by the Italian Association of Aerobiology (A.I.A.), enabled the identification of 26 different pollen types. The pollination graphics show: 1) a late winter period in which pollen from trees appears; 2) a spring period with a prevalence of Gramineae, Fagaceae, Oleaceae andParietaria pollen; 3) a late spring-summer period during which Fagaceae,Pinus and Compositae pollen grains are present; 4) a late summer-autumn and winter period characterized by a drastic reduction of airborne pollen. A study of the calendar reveals, furthermore, that: 1) Gramineae pollen is, amongst those of allergological importance, the most representative; 2)Parietaria is not perennial and is present in modest concentrations; 3)Olea reaches a high peak in June, but not as high as in other southern regions; 3) pollen from arboreal plants is prevalent compared to that of herbaceous plants.     

Effect of exogenous FSH on estrus, ovulation and endogenous hormone release in dairy cows

Fifteen lactating Holstein cows were randomly allotted to receive either 0 mg (group 0), 32 mg (group 1) or 50 mg (group 2) porcine follicle stimulating hormone (FSH-P) injected in 10 fractions at 12 hr intervals beginning on day 9 of the estrous cycle. All cows received 25 mg prostaglandin (PG) on day 11. Jugular blood samples were collected from cows in all groups at 6 hr intervals beginning on day 7 and continuing through expression of estrus. Mean duration to occurrence of estrus and preovulatory LH surge after PG injection was reduced (P<.05) by injection of FSH-P. Mean number of ovulations increased (P<.05) progressively with increased dose of FSH-P. Mean peripheral progesterone declined more uniformly in FSH-P treated cows after PG and increased earlier (P<.05) after estrus in group 2 cows compared to group 0 and 1 cows. Mean plasma estradiol-17beta elevated (P<.05) after PG injection in both FSH-P-treated groups compared to group 0 cows. Both LH and FSH increased (P<.05) for 36 hr after initiation of FSH-P injection in groups 1 and 2, then declined until after PG injection. Peak LH and FSH occurred more uniformly following PG in treated cows. Results indicate that FSH-P increased endogenous gonadotropin release, estradiol-17beta, ovulation rate and reduced duration to estrus and preovulatory gonadotropin release after PG. Injection of 50 mg FSH-P increased plasma estradiol-17beta and ovulation rate compared to injection of 32 mg FSH-P.